Unlock instant, AI-driven research and patent intelligence for your innovation.

Extracellular matrix signaling molecules

a signaling molecule and extracellular matrix technology, applied in the field of extracellular matrix signaling molecules, can solve the problems of affecting the quality of life of afflicted individuals, uncontrolled growth, skeletal defects presenting problems, etc., and achieve the effects of improving tissue grafting, improving neovascularization rate of grafts, and increasing growth

Inactive Publication Date: 2007-06-19
MUNIN
View PDF8 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The present invention provides materials and methods related to extracellular matrix (ECM) signaling molecules. These molecules have important functions in cell adhesion, cell migration, and cell proliferation, as well as in regulating angiogenesis, chondrogenesis, and oncogenesis. The invention includes polynucleotides encoding ECM signaling molecules, polypeptides comprising these molecules, and methods for making and using them. The invention also includes mutations in the cyr61 gene and cells containing these mutations. The technical effects of the invention include improved understanding and manipulation of ECM signaling molecules for various biological activities and applications."

Problems solved by technology

Reentry to the active cell cycle is by necessity tightly regulated, since a breakdown of this control can result in uncontrolled growth, frequently recognized as cancer.
Abnormal elaboration of the programmed development of cells participating in the process of chondrogenesis results in skeletal defects presenting problems that range from cosmetic concerns to life-threatening disorders.
Abnormal progression of angiogenesis or chondrogenesis, as well as mere progression of oncogenesis, substantially impairs the quality of life for afflicted individuals and adds to modern health care costs.
Further characterization of the Cyr61 polypeptide has been hampered by an inability to purify useful quantities of the protein.
Efforts to purify Cyr61 in quantity by overexpression from either eukaryotic or prokaryotic cells typically fail.
One problem associated with attempting to obtain useful quantities of Cyr61 is the reduction in mammalian growth rates induced by overexpression of Cyr61. Another problem with Cyr61 purification is that the cysteine-rich polypeptide, when expressed in bacterial cells using recombinant DNA techniques, is often found in insoluble protein masses.
Therefore, it has been difficult to establish a culture system that mimics in vivo hematopoiesis.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Extracellular matrix signaling molecules
  • Extracellular matrix signaling molecules

Examples

Experimental program
Comparison scheme
Effect test

example 1

Polynucleotide Cloning

[0050]A human cyr61 cDNA was isolated from a human placental cDNA library by probing with the murine cyr61 cDNA sequence using techniques that are standard in the art. See Sambrook et al., incorporated herein by reference. Isolation of the complete murine cyr61 cDNA from a BALB / c 3T3 (ATCC CRL-1658) cDNA library has been described. O'Brien et al., Mol. Cell. Biol. 10:3569–3577 (1990), incorporated herein by reference. The nucleotide and deduced amino acid sequences of murine cyr61 are available from the Genbank database under accession number M32490. The nucleotide sequence of murine cyr61 is presented in SEQ ID NO:1; the murine Cyr61 amino acid sequence is presented in SEQ ID NO:2.

[0051]The human cDNA library was constructed using λgt11 (Promega Corp., Madison, Wis.) as a vector which was transfected into E. coli and plated on LB agar. A murine cDNA expression construct cloned in pGEM-2 (O'Brien et al., [1990]), containing the entire murine cyr61 coding sequen...

example 2

Sequence Analyses

[0067]The nucleotide sequence of murine cyr61 has been described, O'Brien et al. (1990); Latinkic et al., Nucl. Acids Res. 19:3261–3267 (1991), and is set out herein as SEQ ID NO:1.

[0068]The deduced amino acid sequence of murine Cyr61 has been reported, O'Brien et al. (1990), and is set forth in SEQ ID NO:2.

[0069]The nucleotide sequence of the human cyr61 cDNA was determined using the method of Sanger, as described in Sambrook et al. Sequencing templates were generated by constructing a series of nested deletions from a pGEM-2 human cyr61 cDNA clone, as described in Example 1 above. The human cyr61 cDNA sequence is set forth in SEQ ID NO:3. The amino acid sequence of human Cyr61 was deduced from the human cyr61 cDNA sequence and is set forth in SEQ ID NO:4.

[0070]A comparison of the mouse and human Cyr61 sequences, presented in SEQ ID NO:2 and SEQ ID NO:4, respectively, reveals 91% similarity. Both sequences exhibit an N-terminal signal sequence indicative of a proce...

example 3

RNA Analyses

[0073]Polynucleotide probes are useful diagnostic tools for angiogenic, and other, disorders correlated with Cyr61 expression because properly designed probes can reveal the location, and level, of cyr61 gene expression at the transcriptional level. The expression of cyr61, in turn, indicates whether or not genes controlling the process of angiogenesis are being expressed at typical, or expected, levels.

[0074]Using these tools, the mouse cyr61 mRNA expression pattern was determined using an RNase protection technique. O'Brien et al., (1992). In particular, a 289 nucleotide antisense riboprobe was used that would protect 246 nucleotides of the murine cyr61 mRNA (nucleotides 67 to 313 using the numbering of O'Brien et al.) The assays showed levels of cyr61 mRNA in PSA-1 cells (10 μg of total RNA) from either the undifferentiated state or stages 1, 2, and 3 of differentiation (PSA-1 cells undergo three stages of cellular differentiation corresponding to mouse embryonic cell...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
temperatureaaaaaaaaaa
pHaaaaaaaaaa
pHaaaaaaaaaa
Login to View More

Abstract

Polynucleotides encoding mammalian ECM signaling molecules affecting the cell adhesion, migration, and proliferation activities characterizing such complex biological processes as angiogenesis, chondrogenesis, and oncogenesis, are provided. The polynucleotide compositions include DNAs and RNAs comprising part, or all, of an ECM signaling molecule coding sequence, or biological equivalents. Polypeptide compositions are also provided. The polypeptide compositions comprise mammalian ECM signaling molecules, peptide fragments, inhibitory peptides capable of interacting with receptors for ECM signaling molecules, and antibody products recognizing Cyr61. Also provided are methods for producing mammalian ECM signaling molecules. Further provided are methods for using mammalian ECM signaling molecules to screen for, and / or modulate, disorders associated with angiogenesis, chondrogenesis, and oncogenesis; ex vivo methods for using mammalian ECM signaling molecules to prepare blood products are also provided.

Description

[0001]This application is a divisional of U.S. application Ser. No. 09 / 495,448, filed Jan. 31, 2000, now U.S. Pat. No. 6,790,606, which is a continuation-in-part of U.S. application Ser. No. 09 / 142,569, filed Apr. 2, 1999, now U.S. Pat. No. 6,413,735, which is the National Stage of International Application No. PCT / US97 / 04193, filed Mar. 14, 1997, which claims priority to U.S. Provisional Application No. 60 / 013,958, filed Mar. 15, 1996, all of which are incorporated herein by reference.FIELD OF THE INVENTION[0002]The present invention is directed to materials and methods involving extracellular matrix signaling molecules in the form of polypeptides involved in cellular responses to growth factors. More particularly, the invention is directed to Cyr61-, Fisp12-, and CTGF-related polynucleotides, polypeptides, compositions thereof, methods of purifying these polypeptides, and methods of using these polypeptides.BACKGROUND OF THE INVENTION[0003]The growth of mammalian cells is tightly ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(United States)
IPC IPC(8): C12N5/00C12N5/06C12N5/08A61K38/00A61K48/00C07K14/475G01N33/50G01N33/68
CPCC07K14/475G01N33/5008G01N33/5029G01N33/5044G01N33/5055G01N33/5064G01N33/5073G01N33/68A01K2217/05A61K38/00A61K48/00C12N2799/026
Inventor LAU, LESTER F.
Owner MUNIN